title: Ang-2-VEGF-A CrossMab, a novel bispecific human IgG1 antibody blocking VEGF-A and Ang-2 functions simultaneously, mediates potent antitumor, antiangiogenic, and antimetastatic efficacy.
abstract: OBJECTIVE
  VEGF-A blockade has been clinically validated as a treatment for human cancers. Angiopoietin-2 (Ang-2) expression has been shown to function as a key regulator of tumor angiogenesis and metastasis.
  
  
  METHODS
  We have applied the recently developed CrossMab technology for the generation of a bispecific antibody recognizing VEGF-A with one arm based on bevacizumab (Avastin), and the other arm recognizing Ang-2 based on LC06, an Ang-2 selective human IgG1 antibody. The potency of Ang-2-VEGF CrossMab was evaluated alone and in combination with chemotherapy using orthotopic and subcutaneous xenotransplantations, along with metastasis analysis by quantitative real-time Alu-PCR and ex vivo evaluation of vessels, hypoxia, proliferation, and apoptosis. The mechanism of action was further elucidated using Western blotting and ELISA assays.
  
  
  RESULTS
  Ang-2-VEGF-A CrossMab showed potent tumor growth inhibition in a panel of orthotopic and subcutaneous syngeneic mouse tumors and patient or cell line-derived human tumor xenografts, especially at later stages of tumor development. Ang-2-VEGF-A CrossMab treatment led to a strong inhibition of angiogenesis and an enhanced vessel maturation phenotype. Neoadjuvant combination with chemotherapy resulted in complete tumor regression in primary tumor-bearing Ang-2-VEGF-A CrossMab-treated mice. In contrast to Ang-1 inhibition, anti-Ang-2-VEGF-A treatment did not aggravate the adverse effect of anti-VEGF treatment on physiologic vessels. Moreover, treatment with Ang-2-VEGF-A CrossMab resulted in inhibition of hematogenous spread of tumor cells to other organs and reduced micrometastatic growth in the adjuvant setting.
  
  
  CONCLUSIONS
  These data establish Ang-2-VEGF-A CrossMab as a promising antitumor, antiangiogenic, and antimetastatic agent for the treatment of cancer.
authors.affiliation: Authors' Affiliations: Discovery Oncology and Biologics Research, Pharmaceutical Research and Early Development (pRED), Roche Diagnostics GmbH, Penzberg; Department of Vascular Biology and Tumor Angiogenesis, Medical Faculty Mannheim (CBTM), Heidelberg University; Division of Vascular Oncology and Metastasis, German Cancer Research Center Heidelberg (DKFZ-ZMBH Alliance), Heidelberg, Germany; Discovery Oncology, pRED, Roche Glycart AG, Schlieren, Switzerland; Department of Nuclear Medicine, Singapore General Hospital; National Cancer Center, Singapore; Roche Translational Medicine Hub, Singapore; and Roche Products Limited, Welwyn Garden City, United Kingdom.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Kienast
authors.forename: Christian
authors.initials: C
authors.lastname: Klein
authors.forename: Werner
authors.initials: W
authors.lastname: Scheuer
authors.forename: Romi
authors.initials: R
authors.lastname: Raemsch
authors.forename: Erica
authors.initials: E
authors.lastname: Lorenzon
authors.forename: Dirk
authors.initials: D
authors.lastname: Bernicke
authors.forename: Frank
authors.initials: F
authors.lastname: Herting
authors.forename: Sidney
authors.initials: S
authors.lastname: Yu
authors.forename: Huynh Hung
authors.initials: HH
authors.lastname: The
authors.forename: Laurent
authors.initials: L
authors.lastname: Martarello
authors.forename: Christian
authors.initials: C
authors.lastname: Gassner
authors.forename: Kay-Gunnar
authors.initials: KG
authors.lastname: Stubenrauch
authors.forename: Kate
authors.initials: K
authors.lastname: Munro
authors.forename: Hellmut G
authors.initials: HG
authors.lastname: Augustin
authors.forename: Markus
authors.initials: M
authors.lastname: Thomas
mesh_terms: D042702:Angiopoietin-2 / Q000037:antagonists & inhibitors / Q000276:immunology*; D000818:Animals; D018033:Antibodies, Bispecific / Q000008:administration & dosage*; D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage; D000068258:Bevacizumab; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D007074:Immunoglobulin G / Q000008:administration & dosage / Q000276:immunology; D051379:Mice; D009362:Neoplasm Metastasis; D009369:Neoplasms / Q000188:drug therapy* / Q000276:immunology; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000276:immunology; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000276:immunology*
chemical_list: D042702:Angiopoietin-2; D018033:Antibodies, Bispecific; D061067:Antibodies, Monoclonal, Humanized; D007074:Immunoglobulin G; D042461:Vascular Endothelial Growth Factor A; D000068258:Bevacizumab